Schematic operation principle of DSLM

The movie shows a schematic illustration of specimen illumination and fluorescence light detection in the standard DSLM imaging mode. The laser beam illuminates the sample from the side and excites fluorophores along a single line inside the specimen. A pair of laser scanners moves the excitation laser line vertically and horizontally. An optically sectioned image is recorded by rapidly scanning an entire plane in the specimen and detecting the fluorescence at a right angle to the illumination axis. The top left inset shows a zoom-in on the illuminated fraction of the specimen, which is embedded in an agarose cylinder and placed inside the specimen chamber. Supplemental movie from Live Cell Imaging, 2nd ed., Cold Spring Harbor Laboratory Press.